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Quantitation of Trastuzumab and an Antibody to SARS-CoV-2 in Minutes Using Affinity Membranes in 96-Well Plates.
Tan, Hui Yin; Yang, Junyan; Linnes, Jacqueline C; Welch, Christopher J; Bruening, Merlin L.
  • Tan HY; Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, United States.
  • Yang J; Department of Chemical and Biomolecular Engineering, University of Notre Dame, Notre Dame, Indiana 46556, United States.
  • Linnes JC; Weldon School of Bioengineering, Purdue University, West Lafayette, Indiana 47907, United States.
  • Welch CJ; Indiana Consortium for Analytical Science & Engineering (ICASE), 410 W. 10th Street, # 1020H, Indianapolis, Indiana 46202, United States.
  • Bruening ML; Department of Chemistry and Biochemistry, University of Notre Dame, Notre Dame, Indiana 46556, United States.
Anal Chem ; 94(2): 884-891, 2022 01 18.
Article in English | MEDLINE | ID: covidwho-1608171
ABSTRACT
Quantitation of therapeutic monoclonal antibodies (mAbs) in human serum could ensure that patients have adequate levels of mAbs for effective treatment. This research describes the use of affinity, glass-fiber membranes in a 96-well-plate format for rapid (<5 min) quantitation of the therapeutic mAb trastuzumab and a mAb against the SARS-CoV-2 spike protein. Adsorption of a poly(acrylic acid)-containing film in membrane pores and activation of the -COOH groups in the film enable covalent-linking of affinity peptides or proteins to the membrane. Passage of mAb-containing serum through the affinity membrane results in mAb capture within 1 min. Subsequent rinsing, binding of a secondary antibody conjugated to a fluorophore, and a second rinse yield mAb-concentration-dependent fluorescence intensities in the wells. Calibration curves established from analyses on different days have low variability and allow determination of mAb levels in separately prepared samples with an average error <10%, although errors in single-replicate measurements may reach 40%. The assays can occur in diluted serum with physiologically relevant mAb concentrations, as well as in undiluted serum. Thus, the combination of 96-well plates containing affinity membranes, a microplate reader, and a simple vacuum manifold affords convenient mAb quantitation in <5 min.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Experimental Studies / Randomized controlled trials Limits: Humans Language: English Journal: Anal Chem Year: 2022 Document Type: Article Affiliation country: Acs.analchem.1c03654

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Full text: Available Collection: International databases Database: MEDLINE Main subject: SARS-CoV-2 / COVID-19 Type of study: Experimental Studies / Randomized controlled trials Limits: Humans Language: English Journal: Anal Chem Year: 2022 Document Type: Article Affiliation country: Acs.analchem.1c03654